Department of Radiation Medicine, Faculty of Naval Medicine, Second Military Medical University , Shanghai , P. R. China.
Int J Radiat Biol. 2013 Dec;89(12):1003-8. doi: 10.3109/09553002.2013.817704. Epub 2013 Aug 2.
To investigate the potential protective role of molecular hydrogen (H(2)) against (12)C(6+) heavy ion radiation, which is a major hazard for space travel and has been also widely used in heavy ion radiotherapy.
H(2) was dissolved in Roswell Park Memorial Institute (RPMI) 1640 medium under high pressure (0.4 Mpa) to a saturated level by using an apparatus produced by our department. A 2-[6-(4'-hydroxy) phenoxy-3H-xanthen-3-on-9-yl] benzoate (HPF) probe and a 2',7'-Dichlorodihydrofluorescein diacetate (H2DCFH-DA) fluorescent dye were used to measure the intracellular reactive oxygen species (ROS) level. Cell apoptosis were determined by double-staining with Annexin V-fluorescein isothiocyanate (Annexin V-FITC) and propidium iodide (PI) as well as a Hoechst 33342 staining method alternatively. Subsequently, cell cycle analysis was performed using a PI staining method and the expression of apoptotic protein was examined by Western blot.
In this study, we demonstrated H(2) reduced ROS level in Human lymphocyte AHH-1 cells as well as in the radiolysis of water. Our data also showed H(2) attenuated (12)C(6+) radiation- induced cell apoptosis and also alleviated radiation-induced G2/M cell cycle arrest. Heavy ion radiation-induced Caspase 3 activation was also inhibited by H(2) treatment.
In conclusion, these data showed that H(2) attenuated (12)C(6+) radiation-induced cell apoptosis through reducing the ROS level and modulating apoptotic molecules, thus indicating the potential of H(2) as a safe and effective radioprotectant.
研究分子氢（H₂）对（12）C⁶⁺重离子辐射的潜在保护作用，重离子辐射是太空旅行的主要危害，也被广泛应用于重离子放射治疗。
使用本部门研制的装置，在高压（0.4 Mpa）下将 H₂溶解于罗塞韦尔公园纪念研究所（RPMI）1640 培养基中至饱和水平。使用 2-[6-(4'-羟基)苯氧基-3H-呫吨-3-酮]苯甲酸（HPF）探针和 2',7'-二氯二氢荧光素二乙酸酯（H₂DCFH-DA）荧光染料来测量细胞内的活性氧（ROS）水平。通过 Annexin V-异硫氰酸荧光素（Annexin V-FITC）和碘化丙啶（PI）双重染色以及 Hoechst 33342 染色法来测定细胞凋亡。随后，通过 PI 染色法进行细胞周期分析，并通过 Western blot 检测凋亡蛋白的表达。
在这项研究中，我们证明 H₂降低了人淋巴细胞 AHH-1 细胞中的 ROS 水平以及水的辐解产物中的 ROS 水平。我们的数据还表明，H₂减轻了（12）C⁶⁺辐射诱导的细胞凋亡，并缓解了辐射诱导的 G2/M 细胞周期阻滞。H₂处理还抑制了重离子辐射诱导的 Caspase 3 激活。
综上所述，这些数据表明 H₂通过降低 ROS 水平和调节凋亡分子来减轻（12）C⁶⁺辐射诱导的细胞凋亡，这表明 H₂作为一种安全有效的辐射防护剂具有潜在应用价值。
